Pre-Clinical Drug Testing in 2D and 3D Human In Vitro Models of Glioblastoma Incorporating Non-Neoplastic Astrocytes: Tunneling Nano Tubules and Mitochondrial Transfer Modulates Cell Behavior and Therapeutic Respons
Pre-Clinical Drug Testing in 2D and 3D Human In Vitro Models of Glioblastoma Incorporating Non-Neoplastic Astrocytes: Tunneling Nano Tubules and Mitochondrial Transfer Modulates Cell Behavior and Therapeutic Respons
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DOI：
10.3390/ijms20236017
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发表时间：
2019-12-01
影响因子：
5.6
通讯作者：
Pilkington, Geoffrey J.
中科院分区：
文献类型：
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作者：
Civita, Prospero;Leite, Diana M.;Pilkington, Geoffrey J.
星形胶质细胞在胶质母细胞瘤(GBM)微环境中的作用尚不清楚，尤其是在细胞侵袭和耐药性方面。为了评估星形胶质细胞在星形胶质细胞中的这一作用，我们建立了一个全人类2D共培养模型和一个基于透明质酸-明胶的3D水凝胶模型(HyStem(TM)-HP)，该模型含有不同比例的GBM细胞和星形胶质细胞。荧光标记的GBM细胞与星形胶质细胞的接触共培养表明，后者促进了GBM细胞的生长和迁移。值得注意的是，直接接触的非肿瘤性星形胶质细胞的存在，即使在共培养中数量很少，也会在GBM中引起耐药性。最近的研究表明，非肿瘤细胞可以沿着隧道纳米管(TNT)转移线粒体，拯救受损的靶癌细胞。在这些研究中，我们使用2D和3D的GBM和星形胶质细胞的体外共培养模型来探索TNT的形成和线粒体的转移。共培养48h后，胶质纤维酸性蛋白(GFAP)阳性的“反应性”星形胶质细胞形成TNT，而3D透明质酸-明胶水凝胶模型中TNT形成的增加更明显。这项研究表明，在肿瘤微环境中，无论是在2D还是3D体外共培养模型中，人星形胶质细胞都可以与GBM细胞形成TNT连接。我们推测，通过TNT连接传递TNT的非肿瘤性线粒体可能与GBM的药物反应以及增殖和迁移有关。
The role of astrocytes in the glioblastoma (GBM) microenvironment is poorly understood; particularly with regard to cell invasion and drug resistance. To assess this role of astrocytes in GBMs we established an all human 2D co-culture model and a 3D hyaluronic acid-gelatin based hydrogel model (HyStem (TM)-HP) with different ratios of GBM cells to astrocytes. A contact co-culture of fluorescently labelled GBM cells and astrocytes showed that the latter promotes tumour growth and migration of GBM cells. Notably, the presence of non-neoplastic astrocytes in direct contact, even in low amounts in co-culture, elicited drug resistance in GBM. Recent studies showed that non-neoplastic cells can transfer mitochondria along tunneling nanotubes (TNT) and rescue damaged target cancer cells. In these studies, we explored TNT formation and mitochondrial transfer using 2D and 3D in vitro co-culture models of GBM and astrocytes. TNT formation occurs in glial fibrillary acidic protein (GFAP) positive "reactive" astrocytes after 48 h co-culture and the increase of TNT formations was greater in 3D hyaluronic acid-gelatin based hydrogel models. This study shows that human astrocytes in the tumour microenvironment, both in 2D and 3D in vitro co-culture models, could form TNT connections with GBM cells. We postulate that the association on TNT delivery non-neoplastic mitochondria via a TNT connection may be related to GBM drug response as well as proliferation and migration.